Olympus FV1000 confocal microscope


Location: Adrian Building G1.
Contact: Dr Kees Straatman (krs5/7085, 2263).


A guide to start up and shut down the confocal can be found here.
A free viewer to access your images can be installed via the University of Leicester Software Centre. It also can be downloaded from here. Select 'Laser Scanning Microscopes' (for PC only). Contact me (krs5) if you encounter any problems.
You can also open your images using ImageJ (with the Bio-Formats plugin installed) or Fiji.

Available objectives

This system can be used for:
- multi-colour imaging
- single optical sections or 3D stacks
- bleaching (FRAP/FLIP)/photo-activation experiments
- spectral imaging
- multiposition time laps
- tile scans

with additional software:
- Fluorescence Correlation Spectroscopy (FCS)
- Raster Image Correlation Spectroscopy for measurement of diffusion coefficients in live cells is also possible. Please contact krs5.

This microscope is an inverted IX81 motorized microscope equipped with a:
- UV laser diode (405 nm, 50 mW)
- Argon laser, multi-line: 457/488/515 nm (40 mW)
- yellow diode laser (559 nm, 20 mW)
- red diode laser (635 nm, 20 mW)

The system is equipped with 6 detectors:
- 3 photomultiplier tubes (PMTs) for normal imaging. Two of the PMTs are equipped with a galvano grating spectral scanning systems for tunable bandwidth imaging what allows for fluorescence emission detection with variable wavelength and spectral imaging.

- 2 high sensitive detectors (HSDs) for detection of weak signal in the green or red channel. These detectors also have a lower noise level but cannot be used for bright signal. They are not fully integrated in the Olympus software! If unsure how to use these detectors please contact krs5.

- 1 transmitted light detector for brightfield or DIC imaging.

Available dichroic mirrors and barrier filters

Channels

Dichroic Mirror (nm)

Barrier Filters (nm)

PMT 1 SDM490, SDM510, SDM560, SDM640, glass, mirror

Variable Barrier Filter

PMT 2 SDM560, SDM640,glass,
mirror, SDM510-560R

Variable Barrier Filter

PMT 3 glass, mirror, SDM640R BA575-620, BA575-675, BA655-755
HSD 1 SDM560 (fixed)  BA505-540 (fixed)
HSD 2 BA575-675 (fixed)

 

The selection of the laser line is controlled via an Acousto Optical Tunable Filter (AOTF). The use of an AOTF allows excitation intensity balancing within an image, flexible area bleaching e.g. for FRAP/FLIP/Photo-activation experiments and line sequential scanning to avoid cross-talk between channels (multi-tracking). The spectral range of the PMT detectors is appr. 300 nm to 850 nm. The detectors can be used in two modes:

  • (1) conventional photo current detection mode (normal mode)
  • (2) high sensitivity photon counting mode.

 

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